准确的植物亚硫酸氢盐测序。
Accurate sodium bisulfite sequencing in plants.
机构信息
Department of Plant Sciences, University of Cambridge, Downing Street, Cambridge, UK.
出版信息
Epigenetics. 2010 Jan 1;5(1):47-9. doi: 10.4161/epi.5.1.10560. Epub 2010 Jan 8.
Abstract
DNA cytosine methylation is a conserved epigenetic modification frequently correlating with transcriptional silencing in a wide variety of eukaryotic organisms. Sodium bisulfite treatment of DNA converts unmethylated cytosine to uracil, while 5-methylated cytosine is protected. We describe techniques that ensure reliable sequencing data following sodium bisulfite conversion and to avoid common pitfalls such as amplification of unconverted DNA and inclusion of sibling clones.
摘要
DNA 胞嘧啶甲基化是一种保守的表观遗传修饰，常与真核生物中广泛的转录沉默相关。DNA 的亚硫酸氢钠处理将未甲基化的胞嘧啶转化为尿嘧啶，而 5-甲基化的胞嘧啶则受到保护。我们描述了确保亚硫酸氢钠转化后获得可靠测序数据的技术，并避免了常见的陷阱，如未转化 DNA 的扩增和同系克隆的包含。
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